Experiment Research In Vitro And Vivo About Effect Of Adenovirus-mediated HIL-10Gene To Mice With Liver Injury

Objective The experiment were made to study the protection effect ofadenovirus-mediated hIL-10with liver injury.First,in vitro,the experiment was to testwhether adenovirus-mediated hIL-10had anti-inflammatory function to injuredcultured hepatocyte.At the same time, in vivo,the research was to observe transductionefficiency and protection effect to hepatic tissue of adenovirus-mediated hIL-10.Methods At first,adenovirus vector that carries hIL-10was constructed.In vitro,thehepatocyte injury model was induced with CCl4.Adenovirus-mediated hIL-10was putinto hepatocytes in protected group firstly.At last,these cells were cultured24hours.Andthen observe the cell survival probability. In vivo, CCl4was used to induce mice modelswith liver injury..Then, Ringer’s solution containing adenovirus-mediated IL-10wasinjected into mice with liver cirrhosis through caudal vein.Later,ALT,AST and fourindex signs (HA;LN;PCIII;IV-C)of mice models were tested, comparing with controlgroup.At the same time, adenovirus’s transduction efficiency was observed by usingenhanced green fluorescent protein under fluorescence microscope.And expression timeof hIL-10in plasma was tested by Elisa and Western blot methods.Results In vitro,hepatocyte of the mice appeared injured obviously after treated withCCl4(terminal consistency0.5%).And the survival probability dropped to59%to72%.Compared with control group,statistical difference is significant(P<0.05).Whencells are infected with Ad5-hIL10(MOI=1,5,10,50),the survival probability is increasing gradually with the value of MOI. When the value of MOI is50,cell survival probabilityreached92%.In vivo,Then, Ringer’s solution containing adenovirus-mediated hIL-10was injected into mice with liver cirrhosis through caudal vein.After a week,level of theALT and AST level of the mice that treated with hIL-10was obviously lower than micewithout treating.Apart from IV-C，the mice’s other index signs of liver cirrhosis(HA;LN;PCIII) that have been treated are obviously lower than that withouttreating.Pathology results demonstrate that this kind of approach can relieve the damageof CCl4on liver and reduce liver fibrosis.Meanwhile, adenovirus-mediated IL-10genehas high transfection efficiency.The experiment results demonstrate that the cells thatexpresses EGFP takes percent of51.2%.However,the expression time was not long.Elisaand Western blot methods in the research discovered that in blood plasma,hIL-10expression level reached highest in8hours,and then decreased.After one week,hIL-10totally disappeared in plasma. At the same time,another result was found that thismethod of injecting hIL-10was safe,and mortality of these mice was0%.Their survivalstate was better than before.Conclusion Adenovirus-mediated hIL-10has good effect on protecting hepatocyte ofmice with liver injury.And this technique is very safe.Thus,adenovirus-mediated hIL-10gene has a promising future in treating liver injury and liver fibrosis.